Anti-Actin antibody [EPR19353-16] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-Actin antibody [EPR19353-16] - BSA and Azide free (AB251552)

This data was developed using ab216070, the same antibody clone in a different buffer formulation.

Actin was immunoprecipitated from 0.35 mg of NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate with ab216070 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab216070 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1,000 dilution

Lane 1 : NIH/3T3 whole cell lysate, 10 μg (Input).
Lane 2 : ab216070 IP in NIH/3T3 whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab216070 in NIH/3T3 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 3 minutes.

Note : Customer optimization may be required.

All lanes:

Immunoprecipitation - Anti-Actin antibody [EPR19353-16] - Loading Control (<a href='/en-us/products/primary-antibodies/actin-antibody-epr19353-16-loading-control-ab216070'>ab216070</a>)

Predicted band size: 42 kDa

false

• WB

Supplier Data

Western blot - Anti-Actin antibody [EPR19353-16] - BSA and Azide free (AB251552)

This data was developed using ab216070, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Actin antibody [EPR19353-16] - Loading Control (<a href='/en-us/products/primary-antibodies/actin-antibody-epr19353-16-loading-control-ab216070'>ab216070</a>) at 1/1000 dilution

Lane 1:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Lane 2:

HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

Lane 3:

A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 4:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 42 kDa

Observed band size: 42 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-Actin antibody [EPR19353-16] - BSA and Azide free (AB251552)

This data was developed using ab216070, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Actin antibody [EPR19353-16] - Loading Control (<a href='/en-us/products/primary-antibodies/actin-antibody-epr19353-16-loading-control-ab216070'>ab216070</a>) at 1/1000 dilution

Lane 1:

Human skeletal muscle lysate at 10 µg

Lane 2:

Human fetal kidney tissue lysate at 10 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/4000 dilution

Predicted band size: 42 kDa

Observed band size: 42 kDa

false

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-Actin antibody [EPR19353-16] - BSA and Azide free (AB251552)

This data was developed using ab216070, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Human Gamma actin (ACTG1) contains aa2-182 with His-tag. Human Beta actin (ACTB) contains aa1-100 with GST and His tag. Human Gamma-2 actin (ACTG2) contains aa1-200 with His-tag. Human Alpha-cardiac actin (ACTC) contains aa1-200 with His-tag. Human Alpha-actin-2 (ACTA2) contains aa3-253 with His-tag. Human Alpha-actin-1 (ACTA1) contains aa3-200 with His-tag.

All lanes:

Western blot - Anti-Actin antibody [EPR19353-16] - Loading Control (<a href='/en-us/products/primary-antibodies/actin-antibody-epr19353-16-loading-control-ab216070'>ab216070</a>) at 1/1000 dilution

Lane 1:

Human Gamma actin (ACTG1) recombinant protein at 0.01 µg

Lane 2:

Human Beta actin (ACTB) recombinant protein at 0.02 µg

Lane 3:

Human Gamma-2 actin (ACTG2) recombinant protein at 0.02 µg

Lane 4:

Human Alpha-cardiac actin (ACTC) recombinant protein at 0.02 µg

Lane 5:

Human Alpha-actin-2 (ACTA2) recombinant protein at 0.01 µg

Lane 6:

Human Alpha-actin-1 (ACTA1) recombinant protein at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 42 kDa

false

Exposure time: 1min

• WB

Supplier Data

Western blot - Anti-Actin antibody [EPR19353-16] - BSA and Azide free (AB251552)

This data was developed using ab216070, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Actin antibody [EPR19353-16] - Loading Control (<a href='/en-us/products/primary-antibodies/actin-antibody-epr19353-16-loading-control-ab216070'>ab216070</a>) at 1/1000 dilution

Lane 1:

C6 (Rat glial tumor cell line) whole cell lysate at 10 µg

Lane 2:

PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

Lane 3:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 µg

Lane 4:

Mouse brain lysate at 10 µg

Lane 5:

Mouse spleen lysate at 10 µg

Lane 6:

Rat spleen lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 42 kDa

Observed band size: 42 kDa

false

Exposure time: 3min